Canola variety 45H33

ABSTRACT

Provided is a canola variety designated 45H33 and seed, plants and plant parts thereof produced from a cross of inbred varieties. Methods for producing a canola variety comprise crossing canola variety 45H33 with another canola plant. Methods for producing a canola plant containing in its genetic material one or more traits introgressed into 45H33 through backcross conversion and/or transformation, and to the canola seed, plant and plant part produced thereby are described. Canola variety 45H33, the seed, the plant produced from the seed, plant parts and variants, mutants, and minor modifications of canola variety 45H33 are disclosed.

FIELD

The discovery is in the field of Brassica napus breeding (i.e., canolabreeding), specifically relating to the canola variety designated 45H33.

BACKGROUND

The present discovery relates to a novel rapeseed variety designated45H33 which is the result of years of careful breeding and selection.Since such variety is of high quality and possesses a relatively lowlevel of erucic acid in the vegetable oil component and a relatively lowlevel of glucosinolate content in the meal component, it can be termed“canola” in accordance with the terminology commonly used by plantscientists.

The goal of plant breeding is to combine in a single variety or hybridvarious desirable traits. For field crops, these traits may includeresistance to diseases and insects, tolerance to heat and drought,reducing the time to crop maturity, greater yield, and better agronomicquality. With mechanical harvesting of many crops, uniformity of plantcharacteristics such as germination and stand establishment, growthrate, maturity, and plant and pod height, is important. The creation ofnew superior, agronomically sound, and stable high-yielding cultivars ofmany plant types including canola has posed an ongoing challenge toplant breeders. Therefore, there is a continuing need in the field ofagriculture for canola plants having desirable agronomic and industrialcharacteristics.

SUMMARY

A novel Brassica napus variety designated 45H33 is provided. Thisdiscovery thus relates to the seeds of the 45H33 variety, to plants ofthe 45H33 variety, and to methods for producing a canola plant bycrossing the 45H33 variety with itself or another canola plant (whetherby use of male sterility or open pollination), and to methods forproducing a canola plant containing in its genetic material one or moretransgenes, and to transgenic plants produced by that method. Thisdiscovery also relates to canola seeds and plants produced by crossingthe variety 45H33 with another line.

DEFINITIONS

In the description and tables which follow, a number of terms are used.In order to aid in a clear and consistent understanding of thespecification, the following definitions and evaluation criteria areprovided.

Anther Fertility. The ability of a plant to produce pollen; measured bypollen production. 1=sterile, 9=all anthers shedding pollen (vs. PollenFormation which is amount of pollen produced).

Anther Arrangement. The general disposition of the anthers in typicalfully opened flowers is observed.

Chlorophyll Content. The typical chlorophyll content of the mature seedsis determined by using methods recommended by the Western CanadaCanola/Rapeseed Recommending Committee (WCC/RRC). 1=low (less than 8ppm), 2=medium (8 to 15 ppm), 3=high (greater than 15 ppm). Also,chlorophyll could be analyzed using NIR (Near Infrared) spectroscopy aslong as the instrument is calibrated according to the manufacturer'sspecifications.

CMS. Abbreviation for cytoplasmic male sterility.

Cotyledon. A cotyledon is a part of the embryo within the seed of aplant; it is also referred to as a seed leaf. Upon germination, thecotyledon may become the embryonic first leaf of a seedling.

Cotyledon Length. The distance between the indentation at the top of thecotyledon and the point where the width of the petiole is approximately4 mm.

Cotyledon Width. The width at the widest point of the cotyledon when theplant is at the two to three-leaf stage of development. 3=narrow,5=medium, 7=wide.

CV %: Abbreviation for coefficient of variation.

Disease Resistance: Resistance to various diseases is evaluated and isexpressed on a scale of 0=not tested, 1=resistant, 3=moderatelyresistant, 5=moderately susceptible, 7=susceptible, and 9=highlysusceptible.

Erucic Acid Content: The percentage of the fatty acids in the form ofC22:1.as determined by one of the methods recommended by the WCC/RRC,being AOCS Official Method Ce 2-66 Preparation of Methyl esters ofLong-Chain Fatty Acids or AOCS Official Method Ce 1-66 Fatty AcidComposition by Gas Chromatography.

Fatty Acid Content: The typical percentages by weight of fatty acidspresent in the endogenously formed oil of the mature whole dried seedsare determined. During such determination the seeds are crushed and areextracted as fatty acid methyl esters following reaction with methanoland sodium methoxide. Next the resulting ester is analyzed for fattyacid content by gas liquid chromatography using a capillary column whichallows separation on the basis of the degree of unsaturation and fattyacid chain length. This procedure is described in the work of Daun, etal., (1983) J. Amer. Oil Chem. Soc. 60:1751 to 1754 which is hereinincorporated by reference.

Flower Bud Location. A determination is made whether typical buds aredisposed above or below the most recently opened flowers.

Flower Date 50%. (Same as Time to Flowering) The number of days fromplanting until 50% of the plants in a planted area have at least oneopen flower.

Flower Petal Coloration. The coloration of open exposed petals on thefirst day of flowering is observed.

Frost Tolerance (Spring Type Only). The ability of young plants towithstand late spring frosts at a typical growing area is evaluated andis expressed on a scale of 1 (poor) to 5 (excellent).

Gene Silencing. The interruption or suppression of the expression of agene at the level of transcription or translation.

Genotype. Refers to the genetic constitution of a cell or organism.

Glucosinolate Content. The total glucosinolates of seed at 8.5%moisture, as measured by AOCS Official Method AK-1-92 (determination ofglucosinolates content in rapeseed-colza by HPLC), is expressed asmicromoles per gram of defatted, oil-free meal. Capillary gaschromatography of the trimethylsityl derivatives of extracted andpurified desulfoglucosinolates with optimization to obtain optimumindole glucosinolate detection is described in “Procedures of theWestern Canada Canola/Rapeseed Recommending Committee Incorporated forthe Evaluation and Recommendation for Registration of Canola/RapeseedCandidate Cultivars in Western Canada”. Also, glucosinolates could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications.

Grain. Seed produced by the plant or a self or sib of the plant that isintended for food or feed use.

Green Seed. The number of seeds that are distinctly green throughout asdefined by the Canadian Grain Commission. Expressed as a percentage ofseeds tested.

Herbicide Resistance: Resistance to various herbicides when applied atstandard recommended application rates is expressed on a scale of 1(resistant), 2 (tolerant), or 3 (susceptible).

Leaf Anthocyanin Coloration. The presence or absence of leaf anthocyanincoloration, and the degree thereof if present, are observed when theplant has reached the 9- to 11-leaf stage.

Leaf Attachment to Stem. The presence or absence of clasping where theleaf attaches to the stem, and when present the degree thereof, areobserved.

Leaf Attitude. The disposition of typical leaves with respect to thepetiole is observed when at least 6 leaves of the plant are formed.

Leaf Color. The leaf blade coloration is observed when at least sixleaves of the plant are completely developed.

Leaf Glaucosity. The presence or absence of a fine whitish powderycoating on the surface of the leaves, and the degree thereof whenpresent, are observed.

Leaf Length. The length of the leaf blades and petioles are observedwhen at least six leaves of the plant are completely developed.

Leaf Lobes. The fully developed upper stem leaves are observed for thepresence or absence of leaf lobes when at least 6 leaves of the plantare completely developed.

Leaf Margin Indentation. A rating of the depth of the indentations alongthe upper third of the margin of the largest leaf. 1=absent or very weak(very shallow), 3=weak (shallow), 5=medium, 7=strong (deep), 9=verystrong (very deep).

Leaf Margin Hairiness. The leaf margins of the first leaf are observedfor the presence or absence of pubescence, and the degree thereof, whenthe plant is at the two leaf-stage.

Leaf Margin Shape. A visual rating of the indentations along the upperthird of the margin of the largest leaf. 1=undulating, 2=rounded,3=sharp.

Leaf Surface. The leaf surface is observed for the presence or absenceof wrinkles when at least six leaves of the plant are completelydeveloped.

Leaf Tip Reflexion. The presence or absence of bending of typical leaftips and the degree thereof, if present, are observed at the six toeleven leaf-stage.

Leaf Upper Side Hairiness. The upper surfaces of the leaves are observedfor the presence or absence of hairiness, and the degree thereof ifpresent, when at least six leaves of the plant are formed.

Leaf Width. The width of the leaf blades is observed when at least sixleaves of the plant are completely developed.

Locus. A specific location on a chromosome.

Locus Conversion. A locus conversion refers to plants within a varietythat have been modified in a manner that retains the overall genetics ofthe variety and further comprises one or more loci with a specificdesired trait, such as male sterility, insect, disease or herbicideresistance. Examples of single locus conversions include mutant genes,transgenes and native traits finely mapped to a single locus. One ormore locus conversion traits may be introduced into a single canolavariety.

Lodging Resistance. Resistance to lodging at maturity is observed. 1=nottested, 3=poor, 5=fair, 7=good, 9=excellent.

LSD. Abbreviation for least significant difference.

Maturity. The number of days from planting to maturity is observed, withmaturity being defined as the plant stage when pods with seed changecolor, occurring from green to brown or black, on the bottom third ofthe pod-bearing area of the main stem.

NMS. Abbreviation for nuclear male sterility.

Number of Leaf Lobes. The frequency of leaf lobes, when present, isobserved when at least six leaves of the plant are completely developed.

Oil Content: The typical percentage by weight oil present in the maturewhole dried seeds is determined by ISO 10565:1993 Oilseeds Simultaneousdetermination of oil and water—Pulsed NMR method. Also, oil could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications,reference AOCS Procedure Am 1-92 Determination of Oil, Moisture andVolatile Matter, and Protein by Near-Infrared Reflectance.

Pedicel Length. The typical length of the silique stem when mature isobserved. 3=short, 5=medium, 7=long.

Petal Length. The lengths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petal Width. The widths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petiole Length. The length of the petioles is observed, in a lineforming lobed leaves, when at least six leaves of the plant arecompletely developed. 3=short, 5=medium, 7=long.

Plant Height. The overall plant height at the end of flowering isobserved. 3=short, 5=medium, 7=tall.

Ploidy. This refers to the number of chromosomes exhibited by the line,for example diploid or tetraploid.

Pod Anthocyanin Coloration. The presence or absence at maturity ofsilique anthocyanin coloration, and the degree thereof if present, areobserved.

Pod (Silique) Beak Length. The typical length of the silique beak whenmature is observed. 3=short, 5=medium, 7=long.

Pod Habit. The typical manner in which the siliques are borne on theplant at maturity is observed.

Pod (Silique) Length. The typical silique length is observed. 1=short(less than 7 cm), 5=medium (7 to 10 cm), 9=long (greater than 10 cm).

Pod (Silique) Attitude. A visual rating of the angle joining the pedicelto the pod at maturity. 1=erect, 3=semi-erect, 5=horizontal,7=semi-drooping, 9=drooping.

Pod Type. The overall configuration of the silique is observed.

Pod (Silique) Width. The typical pod width when mature is observed.3=narrow (3 mm), 5=medium (4 mm), 7=wide (5 mm).

Pollen Formation. The relative level of pollen formation is observed atthe time of dehiscence.

Protein Content: The typical percentage by weight of protein in the oilfree meal of the mature whole dried seeds is determined by AOCS OfficialMethod Ba 4e-93 Combustion Method for the Determination of CrudeProtein. Also, protein could be analyzed using NIR (Near Infrared)spectroscopy as long as the instrument is calibrated according to themanufacturer's specifications, reference AOCS Procedure Am 1-92Determination of Oil, Moisture and Volatile Matter, and Protein byNear-Infrared Reflectance.

Resistance. The ability of a plant to withstand exposure to an insect,disease, herbicide, or other condition. A resistant plant variety orhybrid will have a level of resistance higher than a comparablewild-type variety or hybrid. “Tolerance” is a term commonly used incrops such as canola, soybean, and sunflower affected by an insect,disease, such as Sclerotinia, herbicide, or other condition and is usedto describe an improved level of field resistance.

Root Anthocyanin Coloration. The presence or absence of anthocyanincoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Anthocyanin Expression. When anthocyanin coloration is present inskin at the top of the root, it further is observed for the exhibitionof a reddish or bluish cast within such coloration when the plant hasreached at least the six-leaf stage.

Root Anthocyanin Streaking. When anthocyanin coloration is present inthe skin at the top of the root, it further is observed for the presenceor absence of streaking within such coloration when the plant hasreached at least the six-leaf stage.

Root Chlorophyll Coloration. The presence or absence of chlorophyllcoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Coloration Below Ground. The coloration of the root skin belowground is observed when the plant has reached at least the six-leafstage.

Root Depth in Soil. The typical root depth is observed when the planthas reached at least the six-leaf stage.

Root Flesh Coloration. The internal coloration of the root flesh isobserved when the plant has reached at least the six-leaf stage.

SE. Abbreviation for standard error.

Seedling Growth Habit. The growth habit of young seedlings is observedfor the presence of a weak or strong rosette character. 1=weak rosette,9=strong rosette.

Seeds Per Pod. The average number of seeds per pod is observed.

Seed Coat Color. The seed coat color of typical mature seeds isobserved. 1=black, 2=brown, 3=tan, 4=yellow, 5=mixed, 6=other.

Seed Coat Mucilage. The presence or absence of mucilage on the seed coatis determined and is expressed on a scale of 1 (absent) to 9 (present).During such determination a petri dish is filled to a depth of 0.3 cm.with water provided at room temperature. Seeds are added to the petridish and are immersed in water where they are allowed to stand for fiveminutes. The contents of the petri dish containing the immersed seedsare then examined under a stereo microscope equipped with transmittedlight. The presence of mucilage and the level thereof is observed as theintensity of a halo surrounding each seed.

Seed Size. The weight in grams of 1,000 typical seeds is determined atmaturity while such seeds exhibit a moisture content of approximately 5to 6 percent by weight.

Shatter Resistance. Resistance to silique shattering is observed at seedmaturity. 1=not tested, 3=poor, 5=fair, 7=good, 9=does not shatter.

SI. Abbreviation for self-incompatible.

Speed of Root Formation. The typical speed of root formation is observedwhen the plant has reached the four to eleven-leaf stage.

SSFS. Abbreviation for Sclerotinia sclerotiorum Field Severity score, arating based on both percentage infection and disease severity.

Stem Anthocyanin Intensity. The presence or absence of leaf anthocyanincoloration and the intensity thereof, if present, are observed when theplant has reached the nine to eleven-leaf stage. 1=absent or very weak,3=weak, 5=medium, 7=strong, 9=very strong.

Stem Lodging at Maturity. A visual rating of a plant's ability to resiststem lodging at maturity. 1=very weak (lodged), 9=very strong (erect).

Time to Flowering. A determination is made of the number of days when atleast 50 percent of the plants have one or more open buds on a terminalraceme in the year of sowing.

Seasonal Type. This refers to whether the new line is considered to beprimarily a Spring or Winter type of canola.

Winter Survival (Winter Type Only). The ability to withstand wintertemperatures at a typical growing area is evaluated and is expressed ona scale of 1 (poor) to 5 (excellent).

DETAILED DESCRIPTION

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant or a genetically identical plant. A plant is sib-pollinated whenindividuals within the same family or line are used for pollination. Aplant is cross-pollinated if the pollen comes from a flower on agenetically different plant from a different family or line. The term“cross-pollination” used herein does not include self-pollination orsib-pollination.

In the practical application of a chosen breeding program, the breederoften initially selects and crosses two or more parental lines, followedby repeated selfing and selection, thereby producing many unique geneticcombinations. The breeder can theoretically generate billions ofdifferent genetic combinations via crossing, selfing and mutagenesis.However, the breeder commonly has no direct control at the cellularlevel of the plant. Therefore, two breeders will never independentlydevelop the same variety having the same canola traits.

In each cycle of evaluation, the plant breeder selects the germplasm toadvance to the next generation. This germplasm is grown under chosengeographical, climatic and soil conditions, and further selections arethen made during and at the end of the growing season. Thecharacteristics of the varieties developed are incapable of predictionin advance. This unpredictability is because the selection occurs inunique environments, with no control at the DNA level (usingconventional breeding procedures), and with millions of differentpossible genetic combinations being generated. A breeder of ordinaryskill cannot predict in advance the final resulting varieties that areto be developed, except possibly in a very gross and general fashion.Even the same breeder is incapable of producing the same variety twiceby using the same original parents and the same selection techniques.This unpredictability commonly results in the expenditure of largeresearch monies and effort to develop a new and superior canola variety.

Canola breeding programs utilize techniques such as mass and recurrentselection, backcrossing, pedigree breeding and haploidy. For a generaldescription of rapeseed and Canola breeding, see, Downey and Rakow,(1987) “Rapeseed and Mustard” In: Principles of Cultivar Development,Fehr, (ed.), pp 437-486; New York; Macmillan and Co.; Thompson, (1983)“Breeding winter oilseed rape Brassica napus”; Advances in AppliedBiology 7:1-104; and Ward, et. al., (1985) Oilseed Rape, Farming PressLtd., Wharfedale Road, Ipswich, Suffolk, each of which is herebyincorporated by reference.

Recurrent selection is used to improve populations of either self- orcross-pollinating Brassica. Through recurrent selection, a geneticallyvariable population of heterozygous individuals is created byintercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, and/or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes.

Breeding programs use backcross breeding to transfer genes for a simplyinherited, highly heritable trait into another line that serves as therecurrent parent. The source of the trait to be transferred is calledthe donor parent. After the initial cross, individual plants possessingthe desired trait of the donor parent are selected and are crossed(backcrossed) to the recurrent parent for several generations. Theresulting plant is expected to have the attributes of the recurrentparent and the desirable trait transferred from the donor parent. Thisapproach has been used for breeding disease resistant phenotypes of manyplant species, and has been used to transfer low erucic acid and lowglucosinolate content into lines and breeding populations of Brassica.

Pedigree breeding and recurrent selection breeding methods are used todevelop varieties from breeding populations. Pedigree breeding startswith the crossing of two genotypes, each of which may have one or moredesirable characteristics that is lacking in the other or whichcomplements the other. If the two original parents do not provide all ofthe desired characteristics, other sources can be included in thebreeding population. In the pedigree method, superior plants are selfedand selected in successive generations. In the succeeding generationsthe heterozygous condition gives way to homogeneous lines as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more generations of selfing and selection arepracticed: F₁ to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅, etc. For example, twoparents that are believed to possess favorable complementary traits arecrossed to produce an F₁. An F₂ population is produced by selfing one orseveral F₁'s or by intercrossing two F₁'s (i.e., sib mating). Selectionof the best individuals may begin in the F₂ population, and beginning inthe F₃ the best individuals in the best families are selected.Replicated testing of families can begin in the F₄ generation to improvethe effectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines commonly are tested forpotential release as new cultivars. Backcrossing may be used inconjunction with pedigree breeding; for example, a combination ofbackcrossing and pedigree breeding with recurrent selection can be usedto incorporate blackleg resistance into certain cultivars of Brassicanapus. Other traits which can be introgressed through backcrossing orgenetic transformation include resistance to Fusarium wilt and clubroot.

Plants that have been self-pollinated and selected for type for manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny. If desired, double-haploidmethods can also be used to extract homogeneous lines. A cross betweentwo different homozygous lines produces a uniform population of hybridplants that may be heterozygous for many gene loci. A cross of twoplants each heterozygous at a number of gene loci will produce apopulation of hybrid plants that differ genetically and will not beuniform.

The choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially, such as F₁ hybrid variety or openpollinated variety. A true breeding homozygous line can also be used asa parental line (inbred line) in a commercial hybrid. If the line isbeing developed as an inbred for use in a hybrid, an appropriatepollination control system should be incorporated in the line.Suitability of an inbred line in a hybrid combination will depend uponthe combining ability (general combining ability or specific combiningability) of the inbred.

Various breeding procedures are also utilized with these breeding andselection methods. The single-seed descent procedure in the strict senserefers to planting a segregating population, harvesting a sample of oneseed per plant, and using the one-seed sample to plant the nextgeneration. When the population has been advanced from the F₂ to thedesired level of inbreeding, the plants from which lines are derivedwill each trace to different F₂ individuals. The number of plants in apopulation declines each generation due to failure of some seeds togerminate or some plants to produce at least one seed. As a result, notall of the F₂ plants originally sampled in the population will berepresented by a progeny when generation advance is completed.

In a multiple-seed procedure, canola breeders commonly harvest one ormore pods from each plant in a population and thresh them together toform a bulk. Part of the bulk is used to plant the next generation andpart is put in reserve. The procedure has been referred to as modifiedsingle-seed descent or the pod-bulk technique. The multiple-seedprocedure has been used to save labor at harvest. It is considerablyfaster to thresh pods with a machine than to remove one seed from eachby hand for the single-seed procedure. The multiple-seed procedure alsomakes it possible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed. If desired,doubled-haploid methods can be used to extract homogeneous lines.

Molecular markers, including techniques such as Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) and Single NucleotidePolymorphisms (SNPs), may be used in plant breeding methods. One use ofmolecular markers is Quantitative Trait Loci (QTL) mapping. QTL mappingis the use of markers which are known to be closely linked to allelesthat have measurable effects on a quantitative trait. Selection in thebreeding process is based upon the accumulation of markers linked to thepositive effecting alleles and/or the elimination of the markers linkedto the negative effecting alleles in the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against themarkers of the donor parent. Using this procedure can minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called Genetic Marker EnhancedSelection or Marker Assisted Selection (MAS).

The production of doubled haploids can also be used for the developmentof inbreds in the breeding program. In Brassica napus, microsporeculture technique is used in producing haploid embryos. The haploidembryos are then regenerated on appropriate media as haploid plantlets,doubling chromosomes of which results in doubled haploid plants. Thiscan be advantageous because the process omits the generations of selfingneeded to obtain a homozygous plant from a heterozygous source.

The development of a canola hybrid in a canola plant breeding programinvolves three steps: (1) the selection of plants from various germplasmpools for initial breeding crosses; (2) the selfing of the selectedplants from the breeding crosses for several generations to produce aseries of inbred lines, which, although different from each other, breedtrue and are highly uniform; and (3) crossing the selected inbred lineswith different inbred lines to produce the hybrids. During theinbreeding process in canola, the vigor of the lines decreases. Vigor isrestored when two different inbred lines are crossed to produce thehybrid. An important consequence of the homozygosity and homogeneity ofthe inbred lines is that the hybrid between a defined pair of inbredswill always be the same. Once the inbreds that give a superior hybridhave been identified, the hybrid seed can be reproduced indefinitely aslong as the homogeneity of the inbred parents is maintained.

Controlling Self-Pollination

Canola varieties are mainly self-pollinated; therefore, self-pollinationof the parental varieties must be controlled to make hybrid developmentfeasible. In developing improved new Brassica hybrid varieties, breedersmay use self-incompatible (SI), cytoplasmic male sterile (CMS) ornuclear male sterile (NMS) Brassica plants as the female parent. Inusing these plants, breeders are attempting to improve the efficiency ofseed production and the quality of the F₁ hybrids and to reduce thebreeding costs. When hybridization is conducted without using SI, CMS orNMS plants, it is more difficult to obtain and isolate the desiredtraits in the progeny (F₁ generation) because the parents are capable ofundergoing both cross-pollination and self-pollination. If one of theparents is a SI, CMS or NMS plant that is incapable of producing pollen,only cross pollination will occur. By eliminating the pollen of oneparental variety in a cross, a plant breeder is assured of obtaininghybrid seed of uniform quality, provided that the parents are of uniformquality and the breeder conducts a single cross.

In one instance, production of F₁ hybrids includes crossing a CMSBrassica female parent with a pollen-producing male Brassica parent. Toreproduce effectively, however, the male parent of the F₁ hybrid musthave a fertility restorer gene (Rf gene). The presence of an Rf genemeans that the F₁ generation will not be completely or partiallysterile, so that either self-pollination or cross pollination may occur.Self-pollination of the F₁ generation to produce several subsequentgenerations is important to ensure that a desired trait is heritable andstable and that a new variety has been isolated.

An example of a Brassica plant which is cytoplasmic male sterile andused for breeding is Ogura (OGU) cytoplasmic male sterile(Pellan-Delourme, et al., 1987). A fertility restorer for Oguracytoplasmic male sterile plants has been transferred from Raphanussativus (radish) to Brassica by Instit. National de Recherche Agricole(INRA) in Rennes, France (Pelletier, et al., 1987). The OGU INRArestorer gene, Rf1 originating from radish, is described in WO 92/05251and in Delourme, et al., (1991). Improved versions of this restorer havebeen developed. For example, see WO98/27806, oilseed brassica containingan improved fertility restorer gene for Ogura cytoplasmic malesterility, which is hereby incorporated by reference.

Other sources and refinements of CMS sterility in canola include thePolima cytoplasmic male sterile plant, as well as those of U.S. Pat. No.5,789,566, DNA sequence imparting cytoplasmic male sterility,mitochondrial genome, nuclear genome, mitochondria and plant containingsaid sequence and process for the preparation of hybrids; U.S. Pat. No.5,973,233 Cytoplasmic male sterility system production canola hybrids;and WO97/02737 Cytoplasmic male sterility system producing canolahybrids; EP Patent Application Number 0 599042A Methods for introducinga fertility restorer gene and for producing F1 hybrids of Brassicaplants thereby; U.S. Pat. No. 6,229,072 Cytoplasmic male sterilitysystem production canola hybrids; U.S. Pat. No. 4,658,085 Hybridizationusing cytoplasmic male sterility, cytoplasmic herbicide tolerance, andherbicide tolerance from nuclear genes; all of which are incorporatedherein for this purpose.

Promising advanced breeding lines commonly are tested and compared toappropriate standards in environments representative of the commercialtarget area(s). The best lines are candidates for new commercial lines;and those still deficient in a few traits may be used as parents toproduce new populations for further selection.

Inbred Development—Female

The female parent is developed by crossing a male sterile version ofvariety NS6945 (A-line) with a maintainer line of variety NS6945(B-line). The A and B lines are genetically alike except the A-linecarries the OGU INRA cytoplasm, while the B-line carries the normal B.napus cytoplasm.

The B-line was developed at Georgetown Research Centre of PioneerHi-Bred Production LP from a cross (NS5102BR×NS5111BR) using pedigreemethod. The last crossing was completed in the summer of 2006 and the F1generation was grown and screened for glyphosate in the greenhouseduring the fall of 2006 and F2 bulk was harvested. The F2 single plantswere planted early in 2007 and screened for glyphosate tolerance witheach F3 harvested individually. These F3s were then evaluated in the2007 Caledon field nursery in for glyphosate tolerance, early maturity,lodging resistance, high oil and protein, total glucosinolates and lowtotal saturated fatty acids, general vigor and uniformity. Remnant seedfrom selected F3s were planted and used as a male parent to create BC0for CMS—A line conversion in the greenhouse during the fall 2007 andsubjected to glyphosate and blackleg screening with F4s harvestedindividually. The F4 single plants were planted early in 2008 andsubjected to glyphosate and blackleg screening with F5s harvestedindividually. These individual F5 plants were grown as rows in Caledon2008 nursery. Selections based on agronomic and quality compositiontraits were made. One of these selections 08SNB066248 was replanted inthe GH with remnant seed and harvested as F6 bulk and was named09SNB03160. This line was advanced 3 more cycles in the greenhouse inbulk during A-line CMS conversion. The F10 was assigned the breeder codeNS6945BR in 2010.

Inbred Development—Male

A male parent or restorer (R line) of variety NS6569 is designatedNS6569MC. The restorer was developed at Georgetown Research Centre ofPioneer Hi-Bred Production LP from a three way cross(NS5706MC×(NS4304MC×Mendel)). The last crossing was completed in 2005.The F1 plants were grown in the greenhouse and were harvested in bulkduring winter months of 2006. The F2 bulks were subsequently grownduring the summer of 2006 and were harvested in bulk. The F3 bulks werethen sent to Chile for single plant bagging. The F4 lines produced onindividual F3 plants from Chile were then evaluated in 2007 nursery inOntario for general vigor, uniformity, homozygosity of restorer gene(RfRf), maturity, oil, protein, total glucosinolates etc. Also, singleplants were selfed and harvested from the selected F4 rows producing F5lines. The F5s were then evaluated at a University of Alberta greenhouseusing an indoor protocol of clubroot screening developed by theUniversity of Alberta. Based on disease score on the large number ofplants for each line, the lines fixed for clubroot resistance wereidentified by December of 2007. Also the same F5 lines were sent toChile for hybrid seed production using NS5102FR as a common female. Eachhybrid combination was contained in a separate cage. The restorer linecarrying genes for resistance to Clubroot were then harvested withhybrid seed from the cages in Chile. One of these hybrids was assigned08N825R as an experimental code. While the hybrid seed was produced,each restorer line was also planted in a separate set of cages forPre-breeder seed increase (F5 to F6). The F6 seed was then harvested andused for multiplying Breeder seed for NS6569MC.

Hybrid Development

45H33 (11N445R) is a fully restored spring Brassica napus hybrid with aglyphosate resistant gene, based on OGU INRA system. It was developed atGeorgetown Research Centre of Pioneer Hi-Bred Production LP (subsidiaryof DuPont Pioneer). It is a single cross hybrid produced by crossing afemale parent (male sterile inbred-A line×maintainer inbred-B line)carrying a glyphosate resistant gene by a restorer—R male line, where Aand B lines are genetically alike except A line carries the OGU INRAcytoplasm, while B line carries the normal B. napus cytoplasm.

A pollination control system and effective transfer of pollen from oneparent to the other offers improved plant breeding and an effectivemethod for producing hybrid canola seed and plants. For example, theOgura cytoplasmic male sterility (CMS) system, developed via protoplastfusion between radish (Raphanus sativus) and rapeseed (Brassica napus),is one of the most frequently used methods of hybrid production. Itprovides stable expression of the male sterility trait (Ogura, 1968,Pelletier, et al., 1983) and an effective nuclear restorer gene (Heyn,1976).

For most traits the true genotypic value may be masked by otherconfounding plant traits or environmental factors. One method foridentifying a superior plant is to observe its performance relative toother experimental plants and to one or more widely grown standardvarieties. If a single observation is inconclusive, replicatedobservations provide a better estimate of the genetic worth.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current varieties. In addition toshowing superior performance, there must be a demand for a new varietythat is compatible with industry standards or which creates a newmarket. The introduction of a new variety commonly will incur additionalcosts to the seed producer, the grower, the processor and the consumer,for special advertising and marketing, altered seed and commercialproduction practices, and new product utilization. The testing precedingrelease of a new variety should take into consideration research anddevelopment costs as well as technical superiority of the final variety.For seed-propagated varieties, it must be feasible to produce seedeasily and economically.

These processes, which lead to the final step of marketing anddistribution, usually take from approximately six to twelve years fromthe time the first cross is made. Therefore, the development of newvarieties is a time-consuming process that requires precise forwardplanning, efficient use of resources, and a minimum of changes indirection.

Further, as a result of the advances in sterility systems, lines aredeveloped that can be used as an open pollinated variety (i.e., apureline cultivar sold to the grower for planting) and/or as a sterileinbred (female) used in the production of F₁ hybrid seed. In the lattercase, favorable combining ability with a restorer (male) would bedesirable. The resulting hybrid seed would then be sold to the growerfor planting.

Combining ability of a line, as well as the performance of the line perse, is a factor in the selection of improved canola lines that may beused as inbreds. Combining ability refers to a line's contribution as aparent when crossed with other lines to form hybrids. The hybrids formedfor the purpose of selecting superior lines are designated test crosses.One way of measuring combining ability is by using breeding values.Breeding values are based on the overall mean of a number of testcrosses. This mean is then adjusted to remove environmental effects andit is adjusted for known genetic relationships among the lines.

Hybrid seed production requires inactivation of pollen produced by thefemale parent. Incomplete inactivation of the pollen provides thepotential for self-pollination. This inadvertently self-pollinated seedmay be unintentionally harvested and packaged with hybrid seed.Similarly, because the male parent is grown next to the female parent inthe field, there is also the potential that the male selfed seed couldbe unintentionally harvested and packaged with the hybrid seed. Once theseed from the hybrid bag is planted, it is possible to identify andselect these self-pollinated plants. These self-pollinated plants willbe genetically equivalent to one of the inbred lines used to produce thehybrid. Though the possibility of inbreds being included in hybrid seedbags exists, the occurrence is rare because much care is taken to avoidsuch inclusions. These self-pollinated plants can be identified andselected by one skilled in the art, through either visual or molecularmethods.

Brassica napus canola plants, absent the use of sterility systems, arerecognized to commonly be self-fertile with approximately 70 to 90percent of the seed normally forming as the result of self-pollination.The percentage of cross pollination may be further enhanced whenpopulations of recognized insect pollinators at a given growing site aregreater. Thus open pollination is often used in commercial canolaproduction.

Since canola variety 45H33 is a hybrid produced from substantiallyhomogeneous parents, it can be reproduced by planting seeds of suchparents, growing the resulting canola plants under controlledpollination conditions with adequate isolation so that cross-pollinationoccurs between the parents, and harvesting the resulting hybrid seedusing conventional agronomic practices.

Locus Conversions of Canola Variety 45H33

45H33 represents a new base genetic line into which a new locus or traitmay be introduced. Direct transformation and backcrossing represent twoimportant methods that can be used to accomplish such an introgression.The term locus conversion is used to designate the product of such anintrogression.

To select and develop a superior hybrid, it is necessary to identify andselect genetically unique individuals that occur in a segregatingpopulation. The segregating population is the result of a combination ofcrossover events plus the independent assortment of specificcombinations of alleles at many gene loci that results in specific andunique genotypes. Once such a variety is developed its value to societyis substantial since it is important to advance the germplasm base as awhole in order to maintain or improve traits such as yield, diseaseresistance, pest resistance and plant performance in extreme weatherconditions. Locus conversions are routinely used to add or modify one ora few traits of such a line and this further enhances its value andusefulness to society.

Backcrossing can be used to improve inbred varieties and a hybridvariety which is made using those inbreds. Backcrossing can be used totransfer a specific desirable trait from one variety, the donor parent,to an inbred called the recurrent parent which has overall goodagronomic characteristics yet that lacks the desirable trait. Thistransfer of the desirable trait into an inbred with overall goodagronomic characteristics can be accomplished by first crossing arecurrent parent to a donor parent (non-recurrent parent). The progenyof this cross is then mated back to the recurrent parent followed byselection in the resultant progeny for the desired trait to betransferred from the non-recurrent parent.

Traits may be used by those of ordinary skill in the art to characterizeprogeny. Traits are commonly evaluated at a significance level, such asa 1%, 5% or 10% significance level, when measured in plants grown in thesame environmental conditions. For example, a locus conversion of 45H33may be characterized as having essentially the same phenotypic traits as45H33. The traits used for comparison may be those traits shown in anyof Tables 1-6. Molecular markers can also be used during the breedingprocess for the selection of qualitative traits. For example, markerscan be used to select plants that contain the alleles of interest duringa backcrossing breeding program. The markers can also be used to selectfor the genome of the recurrent parent and against the genome of thedonor parent. Using this procedure can minimize the amount of genomefrom the donor parent that remains in the selected plants.

A locus conversion of 45H33 will retain the genetic integrity of 45H33.A locus conversion of 45H33 will comprise at least 92%, 93%, 94%, 95%,96%, 97%, 98% or 99% of the base genetics of 45H33. For example, a locusconversion of 45H33 can be developed when DNA sequences are introducedthrough backcrossing (Hallauer et al., 1988), with a parent of 45H33utilized as the recurrent parent. Both naturally occurring andtransgenic DNA sequences may be introduced through backcrossingtechniques. A backcross conversion may produce a plant with a locusconversion in at least one or more backcrosses, including at least 2crosses, at least 3 crosses, at least 4 crosses, at least 5 crosses andthe like. Molecular marker assisted breeding or selection may beutilized to reduce the number of backcrosses necessary to achieve thebackcross conversion. For example, see Openshaw, S. J. et al.,Marker-assisted Selection in Backcross Breeding. In: ProceedingsSymposium of the Analysis of Molecular Data, August 1994, Crop ScienceSociety of America, Corvallis, Oreg., where it is demonstrated that abackcross conversion can be made in as few as two backcrosses.

Uses of Canola

Currently Brassica napus canola is being recognized as an increasinglyimportant oilseed crop and a source of meal in many parts of the world.The oil as removed from the seeds commonly contains a lesserconcentration of endogenously formed saturated fatty acids than othervegetable oils and is well suited for use in the production of salad oilor other food products or in cooking or frying applications. The oilalso finds utility in industrial applications. Additionally, the mealcomponent of the seeds can be used as a nutritious protein concentratefor livestock.

Canola oil has the lowest level of saturated fatty acids of allvegetable oils. “Canola” refers to rapeseed (Brassica) which (1) has anerucic acid (C_(22:1)) content of at most 2 percent by weight based onthe total fatty acid content of a seed, preferably at most 0.5 percentby weight and most preferably essentially 0 percent by weight; and (2)produces, after crushing, an air-dried meal containing less than 30micromoles (μmol) glucosinolates per gram of defatted (oil-free) meal.These types of rapeseed are distinguished by their edibility incomparison to more traditional varieties of the species.

Disease—Sclerotinia

Sclerotinia infects over 100 species of plants, including numerouseconomically important crops such as Brassica species, sunflowers, drybeans, soybeans, field peas, lentils, lettuce, and potatoes (Boland andHall, 1994). Sclerotinia sclerotiorum is responsible for over 99% ofSclerotinia disease, while Sclerotinia minor produces less than 1% ofthe disease. Sclerotinia produces sclerotia, irregularly-shaped, darkoverwintering bodies, which can endure in soil for four to five years.The sclerotia can germinate carpogenically or myceliogenically,depending on the environmental conditions and crop canopies. The twotypes of germination cause two distinct types of diseases. Sclerotiathat germinate carpogenically produce apothecia and ascospores thatinfect above-ground tissues, resulting in stem blight, stalk rot, headrot, pod rot, white mold and blossom blight of plants. Sclerotia thatgerminate myceliogenically produce mycelia that infect root tissues,causing crown rot, root rot and basal stalk rot.

Sclerotinia causes Sclerotinia stem rot, also known as white mold, inBrassica, including canola. Canola is a type of Brassica having a lowlevel of glucosinolates and erucic acid in the seed. The sclerotiagerminate carpogenically in the summer, producing apothecia. Theapothecia release wind-borne ascospores that travel up to one kilometer.The disease is favoured by moist soil conditions (at least 10 days at ornear field capacity) and temperatures of 15-25° C., prior to and duringcanola flowering. The spores cannot infect leaves and stems directly;they must first land on flowers, fallen petals, and pollen on the stemsand leaves. Petal age affects the efficiency of infection, with olderpetals more likely to result in infection (Heran, et al., 1999). Thefungal spores use the flower parts as a food source as they germinateand infect the plant.

The severity of Sclerotinia in Brassica is variable, and is dependent onthe time of infection and climatic conditions (Heran, et al., 1999). Thedisease is favored by cool temperatures and prolonged periods ofprecipitation. Temperatures between 20 and 25° C. and relativehumidities of greater than 80% are required for optimal plant infection(Heran, et al., 1999). Losses ranging from 5 to 100% have been reportedfor individual fields (Manitoba Agriculture, Food and Rural Initiatives,2004). On average, yield losses are estimated to be 0.4 to 0.5 times theSclerotinia sclerotiorum Field Severity score, a rating based on bothpercentage infection and disease severity. More information is providedherein at Example 4. For example, if a field has 20% infection (20/100plants infected), then the yield loss would be about 10% provided plantsare dying prematurely due to the infection of the main stem (rating5-SSFS=20%). If the plants are affected much less (rating 1-SSFS=4%),yield loss is reduced accordingly. Further, Sclerotinia can cause heavylosses in wet swaths. Sclerotinia sclerotiorum caused economic losses tocanola growers in Minnesota and North Dakota of 17.3, 20.8, and 16.8million dollars in 1999, 2000 and 2001, respectively (Bradley, et al.2006). In Canada, this disease is extremely important in SouthernManitoba, parts of South Central Alberta and also in Eastern areas ofSaskatchewan. Since weather plays an important role in development ofthis disease, its occurrence is irregular and unpredictable. Certainreports estimate about 0.8 to 1.3 million acres of canola being sprayedwith fungicide in Southern Manitoba annually. The fungicide applicationcosts about $25 per acre, which represents a significant cost for canolaproducers. Moreover, producers may decide to apply fungicide based onthe weather forecast, while later changes in the weather patterndiscourage disease development, resulting in wasted product, time, andfuel. Creation of Sclerotinia tolerant canola cultivars has been animportant goal for many of the Canadian canola breeding organizations.

The symptoms of Sclerotinia infection usually develop several weeksafter flowering begins. The plants develop pale-grey to white lesions,at or above the soil line and on upper branches and pods. The infectionsoften develop where the leaf and the stem join because the infectedpetals lodge there. Once plants are infected, the mold continues to growinto the stem and invade healthy tissue. Infected stems appear bleachedand tend to shred. Hard black fungal sclerotia develop within theinfected stems, branches, or pods. Plants infected at flowering producelittle or no seed. Plants with girdled stems wilt and ripen prematurely.Severely infected crops frequently lodge, shatter at swathing, and makeswathing more time consuming. Infections can occur in all above-groundplant parts, especially in dense or lodged stands, where plant-to-plantcontact facilitates the spread of infection. New sclerotia carry thedisease over to the next season.

Conventional methods for control of Sclerotinia diseases include (a)chemical control, (b) disease resistance and (c) cultural control, eachof which is described below.

(a) Fungicides such as benomyl, vinclozolin and iprodione remain themain method of control of Sclerotinia disease (Morall, et al., 1985; Tu,1983). Recently, additional fungicidal formulations have been developedfor use against Sclerotinia, including azoxystrobin, prothioconazole,and boscalid. (Johnson, 2005) However, use of fungicide is expensive andcan be harmful to the user and environment. Further, resistance to somefungicides has occurred due to repeated use.

(b) In certain cultivars of bean, safflower, sunflower and soybean, someprogress has been made in developing partial (incomplete) resistance.Partial resistance is often referred to as tolerance. However, successin developing partial resistance has been very limited, probably becausepartial physiological resistance is a multigene trait as demonstrated inbean (Fuller, et al., 1984). In addition to partial physiologicalresistance, some progress has been made to breed for morphologicaltraits to avoid Sclerotinia infection, such as upright growth habit,lodging resistance and narrow canopy. For example, bean plants withpartial physiological resistance and with an upright stature, narrowcanopy and indeterminate growth habit were best able to avoidSclerotinia (Saindon, et al., 1993). Early maturing cultivars ofsafflower showed good field resistance to Sclerotinia. Finally, insoybean, cultivar characteristics such as height, early maturity andgreat lodging resistance result in less disease, primarily because of areduction of favorable microclimate conditions for the disease. (Bolandand Hall, 1987; Buzzell, et al. 1993)

(c) Cultural practices, such as using pathogen-free or fungicide-treatedseed, increasing row spacing, decreasing seeding rate to reducesecondary spread of the disease, and burying sclerotia to preventcarpogenic germination, may reduce Sclerotinia disease but noteffectively control the disease.

All Canadian canola genotypes are susceptible to Sclerotinia stem rot(Manitoba Agriculture, Food and Rural Initiatives, 2004). This includesall known spring petalled genotypes of canola quality. There is also noresistance to Sclerotinia in Australian canola varieties.(Hind-Lanoiselet, et al. 2004). Some varieties with certainmorphological traits are better able to withstand Sclerotinia infection.For example, Polish varieties (Brassica rapa) have lighter canopies andseem to have much lower infection levels. In addition, petal-lessvarieties (apetalous varieties) avoid Sclerotinia infection to a greaterextent (Okuyama, et al., 1995; Fu, 1990). Other examples ofmorphological traits which confer a degree of reduced fieldsusceptibility in Brassica genotypes include increased standability,reduced petal retention, branching (less compact and/or higher), andearly leaf abscission. Jurke and Fernando, (2003) screened eleven canolagenotypes for Sclerotinia disease incidence. Significant variation indisease incidence was explained by plant morphology, and the differencein petal retention was identified as the most important factor. However,these morphological traits alone do not confer resistance toSclerotinia, and all canola products in Canada are consideredsusceptible to Sclerotinia.

Winter canola genotypes are also susceptible to Sclerotinia. In Germany,for example, no Sclerotinia-resistant varieties are available. (Specht,2005) The widely-grown German variety Express is considered susceptibleto moderately susceptible and belongs to the group of less susceptiblevarieties/hybrids.

Spraying with fungicide is the only means of controlling Sclerotinia incanola crops grown under disease-favorable conditions at flowering.Typical fungicides used for controlling Sclerotinia on Brassica includeRovral™/Proline™ from Bayer and Ronilan™/Lance™ from BASF. The activeingredient in Lance™ is Boscalid, and it is marketed as Endura™ in theUnited States. The fungicide should be applied before symptoms of stemrot are visible and usually at the 20-30% bloom stage of the crop. Ifinfection is already evident, there is no use in applying fungicide asit is too late to have an effect. Accordingly, growers must assess theirfields for disease risk to decide whether to apply a fungicide. This canbe done by using a government provided checklist or by using a petaltesting kit. Either method is cumbersome and prone to errors.(Hind-Lanoiselet, 2004; Johnson, 2005)

Numerous efforts have been made to develop Sclerotinia resistantBrassica plants. Built-in resistance would be more convenient,economical, and environmentally-friendly than controlling Sclerotinia byapplication of fungicides. Since the trait is polygenic it would bestable and not prone to loss of efficacy, as fungicides may be.Characteristics of 45H33

Homogenous and reproducible canola hybrids are useful for the productionof a commercial crop on a reliable basis. There are a number ofanalytical methods available to determine the phenotypic stability of acanola hybrid.

The oldest and most traditional method of analysis is the observation ofphenotypic traits. The data are usually collected in field experimentsover the life of the canola plants to be examined. Phenotypiccharacteristics most often are observed for traits associated with seedyield, seed oil content, seed protein content, fatty acid composition ofoil, glucosinolate content of meal, growth habit, lodging resistance,plant height, shatter resistance, etc.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. A plant's genotype can be used to identify plants of thesame variety or a related variety. For example, the genotype can be usedto determine the pedigree of a plant. There are many laboratory-basedtechniques available for the analysis, comparison and characterizationof plant genotype; among these are Isozyme Electrophoresis, RestrictionFragment Length Polymorphisms (RFLPs), Randomly Amplified PolymorphicDNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), SimpleSequence Repeats (SSRs) which are also referred to as Microsatellites,and Single Nucleotide Polymorphisms (SNPs).

The variety of the present discovery has shown uniformity and stabilityfor all traits, as described in the following variety descriptioninformation. The variety has been increased with continued observationfor uniformity.

45H33 is a medium maturing, Clubroot resistant, high yielding glyphosateresistant Brassica napus canola hybrid having resistant (R) rating forblackleg and resistant (R) rating for Fusarium wilt. Its oil content is0.2% lower than WCC/RRC checks. Its protein is slightly higher than meanof the checks and chlorophyll is similar to the checks.

Table 1 provides data on morphological, agronomic, and quality traitsfor 45H33 and canola variety 45H29. When preparing the detailedphenotypic information that follows, plants of the new 45H33 varietywere observed while being grown using conventional agronomic practices.For comparative purposes, canola plants of canola varieties, 45H29 wassimilarly grown in a replicated experiment.

Observations were recorded on various morphological traits for thehybrid 45H33 and comparative check cultivars. (See Table 1).

Hybrid 45H33 can be advantageously used in accordance with the breedingmethods described herein and those known in the art to produce hybridsand other progeny plants retaining desired trait combinations of 45H33.This discovery is thus also directed to methods for producing a canolaplant by crossing a first parent canola plant with a second parentcanola plant wherein either the first or second parent canola plant iscanola variety 45H33. Further, both first and second parent canolaplants can come from the canola variety 45H33. Either the first or thesecond parent plant may be male sterile.

Still further, this discovery also is directed to methods for producinga 45H33-derived canola plant by crossing canola variety 45H33 with asecond canola plant and growing the progeny seed, and repeating thecrossing and growing steps with the canola 45H33-derived plant from 1 to2 times, 1 to 3 times, 1 to 4 times, or 1 to 5 times. Thus, any suchmethods using the canola variety 45H33 are part of this discovery: openpollination, selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using canola variety45H33 as a parent are within the scope of this discovery, includingplants derived from canola variety 45H33. This includes canola linesderived from 45H33 which include components for either male sterility orfor restoration of fertility. Advantageously, the canola variety is usedin crosses with other, different, canola plants to produce firstgeneration (F₁) canola hybrid seeds and plants with superiorcharacteristics.

The discovery also includes a single-gene conversion of 45H33. Asingle-gene conversion occurs when DNA sequences are introduced throughtraditional (non-transformation) breeding techniques, such asbackcrossing. DNA sequences, whether naturally occurring or transgenes,may be introduced using these traditional breeding techniques. Desiredtraits transferred through this process include, but are not limited to,fertility restoration, fatty acid profile modification, othernutritional enhancements, industrial enhancements, disease resistance,insect resistance, herbicide resistance and yield enhancements. Thetrait of interest is transferred from the donor parent to the recurrentparent, in this case, the canola plant disclosed herein. Single-genetraits may result from the transfer of either a dominant allele or arecessive allele. Selection of progeny containing the trait of interestis done by direct selection for a trait associated with a dominantallele. Selection of progeny for a trait that is transferred via arecessive allele will require growing and selfing the first backcross todetermine which plants carry the recessive alleles. Recessive traits mayrequire additional progeny testing in successive backcross generationsto determine the presence of the gene of interest.

It should be understood that the canola variety of the discovery can,through routine manipulation by cytoplasmic genes, nuclear genes, orother factors, be produced in a male-sterile or restorer form asdescribed in the references discussed earlier. Such embodiments are alsowithin the scope of the present claims. Canola variety 45H33 can bemanipulated to be male sterile by any of a number of methods known inthe art, including by the use of mechanical methods, chemical methods,self-incompatibility (SI), cytoplasmic male sterility (CMS) (eitherOgura or another system), or nuclear male sterility (NMS). The term“manipulated to be male sterile” refers to the use of any availabletechniques to produce a male sterile version of canola variety 45H33.The male sterility may be either partial or complete male sterility.This discovery is also directed to F1 hybrid seed and plants produced bythe use of Canola variety 45H33. Canola variety 45H33 can also furthercomprise a component for fertility restoration of a male sterile plant,such as an Rf restorer gene. In this case, canola variety 45H33 couldthen be used as the male plant in hybrid seed production.

This discovery is also directed to the use of 45H33 in tissue culture.As used herein, the term plant includes plant protoplasts, plant celltissue cultures from which canola plants can be regenerated, plantcalli, plant clumps, and plant cells that are intact in plants or partsof plants, such as embryos, pollen, ovules, seeds, flowers, kernels,ears, cobs, leaves, husks, stalks, roots, root tips, anthers, silk andthe like. Pauls, et al., (2006) (Canadian J of Botany 84(4):668-678)confirmed that tissue culture as well as microspore culture forregeneration of canola plants can be accomplished successfully. Chuong,et al., (1985) “A Simple Culture Method for Brassica HypocotylProtoplasts”, Plant Cell Reports 4:4-6; Barsby, et al., (Spring 1996) “ARapid and Efficient Alternative Procedure for the Regeneration of Plantsfrom Hypocotyl Protoplasts of Brassica napus”, Plant Cell Reports;Kartha, et al., (1974) “In vitro Plant Formation from Stem Explants ofRape”, Physiol. Plant 31:217-220; Narasimhulu, et al., (Spring 1988)“Species Specific Shoot Regeneration Response of Cotyledonary Explantsof Brassicas”, Plant Cell Reports; Swanson, (1990) “Microspore Culturein Brassica”, Methods in Molecular Biology 6(17):159; “Cell Culturetechniques and Canola improvement” J. Am. Oil Chem. Soc. 66(4):455-56(1989). Thus, it is clear from the literature that the state of the artis such that these methods of obtaining plants are, and were,“conventional” in the sense that they are routinely used and have a veryhigh rate of success.

The utility of canola variety 45H33 also extends to crosses with otherspecies. Commonly, suitable species will be of the family Brassicae.

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modified versions of native or endogenous geneticelements in order to alter the traits of a plant in a specific manner.Any DNA sequences, whether from a different species, or from the samespecies that are inserted into the genome using transformation arereferred to herein collectively as “transgenes”. Over the last fifteento twenty years several methods for producing transgenic plants havebeen developed, and the present discovery, in particular embodiments,also relates to transformed versions of the claimed canola variety45H33.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,and Genetic Transformation for the improvement of Canola World Conf,Biotechnol. Fats and Oils Ind. 43-46 (1988). In addition, expressionvectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber, et al., “Vectors for Plant Transformation” in Methodsin Plant Molecular Biology and Biotechnology, Glick and Thompson, Eds.(CRC Press, Inc., Boca Raton, 1993) pages 89-119.

The most prevalent types of plant transformation involve theconstruction of an expression vector. Such a vector comprises a DNAsequence that contains a gene under the control of or operatively linkedto a regulatory element, for example a promoter. The vector may containone or more genes and one or more regulatory elements.

A genetic trait which has been engineered into a particular canola plantusing transformation techniques could be moved into another line usingtraditional breeding techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove a transgene from a transformed canola plant to an elite inbred lineand the resulting progeny would comprise a transgene. Also, if an inbredline was used for the transformation then the transgenic plants could becrossed to a different line in order to produce a transgenic hybridcanola plant. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context. Variousgenetic elements can be introduced into the plant genome usingtransformation. These elements include but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences. See,U.S. Pat. No. 6,222,101 which is herein incorporated by reference.

With transgenic plants according to the present discovery, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, (1981) Anal. Biochem.114:92-96.

A genetic map can be generated, primarily via conventional RestrictionFragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR)analysis, Simple Sequence Repeats (SSR), and Single NucleotidePolymorphisms (SNPs), which identifies the approximate chromosomallocation of the integrated DNA molecule coding for the foreign protein.For exemplary methodologies in this regard, see, Glick and Thompson,METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284 (CRC Press,Boca Raton, 1993). Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons would involvehybridizations, RFLP, PCR, SSR, SNP, and sequencing, all of which areconventional techniques.

Likewise, by means of the present discovery, plants can be geneticallyengineered to express various phenotypes of agronomic interest.Exemplary transgenes implicated in this regard include, but are notlimited to, those categorized below.

1. Genes that confer resistance to pests or disease and that encode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones, et al., (1994) Science 266:789(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin, et al., (1993) Science 262:1432 (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinos,et al., (1994) Cell 78:1089 (Arabidopsis RSP2 gene for resistance toPseudomonas syringae); McDowell and Woffenden, (2003) Trends Biotechnol.21(4):178-83 and Toyoda, et al., (2002) Transgenic Res. 11(6):567-82. Aplant resistant to a disease is one that is more resistant to a pathogenas compared to the wild type plant.

(B) A gene conferring resistance to fungal pathogens, such as oxalateoxidase or oxalate decarboxylase (Zhou, et al., (1998) Pl. Physiol.117(1):33-41).

(C) A Bacillus thuringiensis (Bt) protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser, et al.,(1986) Gene 48:109, who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Manassas, Va.), for example, under ATCC Accession Numbers.40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensistransgenes being genetically engineered are given in the followingpatents and patent applications and hereby are incorporated by referencefor this purpose: U.S. Pat. Nos. 5,188,960; 5,689,052; 5,880,275; WO91/114778; WO 99/31248; WO 01/12731; WO 99/24581; WO 97/40162 and U.S.application Ser. Nos. 10/032,717; 10/414,637; and Ser. No. 10/606,320.

(D) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock, et al., (1990) Nature 344:458, of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(E) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, see the disclosures ofRegan, (1994) J. Biol. Chem. 269:9 (expression cloning yields DNA codingfor insect diuretic hormone receptor) and Pratt, et al., (1989) Biochem.Biophys. Res. Comm. 163:1243 (an allostatin is identified in Diplopterapuntata); Chattopadhyay, et al., (2004) Critical Reviews in Microbiology30(1):33-54 2004; Zjawiony, (2004) J Nat Prod 67(2):300-310; Carlini andGrossi-de-Sa, (2002) Toxicon 40(11):1515-1539; Ussuf, et al., (2001)Curr Sci. 80(7):847-853 and Vasconcelos and Oliveira, (2004) Toxicon44(4):385-403. See also, U.S. Pat. No. 5,266,317 to Tomalski, et al.,who disclose genes encoding insect-specific, paralytic neurotoxins.

(F) An enzyme responsible for a hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(G) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTApplication Number WO 93/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase gene. DNA molecules whichcontain chitinase-encoding sequences can be obtained, for example, fromthe ATCC under Accession Numbers 39637 and 67152. See also, Kramer, etal., (1993) Insect Biochem. Molec. Biol. 23:691, who teach thenucleotide sequence of a cDNA encoding tobacco hookworm chitinase, andKawalleck et al., (1993) Plant Molec. Biol. 21:673, who provide thenucleotide sequence of the parsley ubi4-2 polyubiquitin gene, U.S.patent application Ser. Nos. 10/389,432, 10/692,367 and U.S. Pat. No.6,563,020.

(H) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella, et al., (1994) Plant Molec. Biol. 24:757, ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess,et al., (1994) Plant Physiol. 104:1467, who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(I) A hydrophobic moment peptide. See, PCT Application Number WO95/16776and U.S. Pat. No. 5,580,852 (disclosure of peptide derivatives ofTachyplesin which inhibit fungal plant pathogens) and PCT ApplicationNumber WO95/18855 and U.S. Pat. No. 5,607,914 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference for this purpose.

(J) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes, et al., (1993) Plant Sci. 89:43,of heterologous expression of a cecropin-beta lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

(K) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy, et al., (1990) Ann. Rev.Phytopathol. 28:451. Coat protein-mediated resistance has been conferredupon transformed plants against alfalfa mosaic virus, cucumber mosaicvirus, tobacco streak virus, potato virus X, potato virus Y, tobaccoetch virus, tobacco rattle virus and tobacco mosaic virus. Id.

(L) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor, et al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(M) A virus-specific antibody. See, for example, Tavladoraki, et al.,(1993) Nature 366:469, who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(N) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient release bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See, Lamb,et al., (1992) Bio/Technology 10:1436. The cloning and characterizationof a gene which encodes a bean endopolygalacturonase-inhibiting proteinis described by Toubart, et al., (1992) Plant J. 2:367.

(O) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann, et al., (1992) Bio/Technology 10:305, have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(P) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, (1995) Current Biology5(2):128-131, Pieterse and Van Loon, (2004) Curr. Opin. Plant Bio7(4):456-64 and Somssich, (2003) Cell 113(7):815-6.

(Q) Antifungal genes (Cornelissen and Melchers, (1993) Pl. Physiol.101:709-712 and Parijs, et al., (1991) Planta 183:258-264 and Bushnell,et al., (1998) Can. J. of Plant Path. 20(2):137-149. Also see, U.S.patent application Ser. No. 09/950,933.

(R) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see, U.S. Pat. No. 5,792,931.

(S) Cystatin and cysteine proteinase inhibitors. See, U.S. patentapplication Ser. No. 10/947,979.

(T) Defensin genes. See, WO03/000863 and U.S. patent application Ser.No. 10/178,213.

(U) Genes that confer resistance to Phytophthora Root Rot, such as theBrassica equivalents of the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d,Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6,Rps 7 and other Rps genes. See, for example, Shoemaker, et al, (1995)Phytophthora Root Rot Resistance Gene Mapping in Soybean, Plant GenomeIV Conference, San Diego, Calif.

2. Genes that confer resistance to a herbicide, for example:

(A) A herbicide that inhibits the growing point or meristem, such as animidazalinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee, etal., (1988) EMBO J. 7:1241, and Miki, et al., (1990) Theor. Appl. Genet.80:449, respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659;5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;5,928,937 and 5,378,824; and international publication WO 96/33270,which are incorporated herein by reference for this purpose.

(B) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase, PAT) and Streptomyceshygroscopicus phosphinothricin-acetyl transferase, bar, genes), andpyridinoxy or phenoxy propionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah, et al., which discloses the nucleotide sequence of a form of EPSPwhich can confer glyphosate resistance. See also, U.S. Pat. No.7,405,074, and related applications, which disclose compositions andmeans for providing glyphosate resistance. U.S. Pat. No. 5,627,061 toBarry, et al., also describes genes encoding EPSPS enzymes. See also,U.S. Pat. Nos. 6,566,587; 6,338,961; 6,248,876 B1; 6,040,497; 5,804,425;5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835;5,866,775; 6,225,114 B1; 6,130,366; 5,310,667; 4,535,060; 4,769,061;5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and 5,491,288; andinternational publications EP1173580; WO 01/66704; EP1173581 andEP1173582, which are incorporated herein by reference for this purpose.A DNA molecule encoding a mutant aroA gene can be obtained under ATCCAccession Number 39256, and the nucleotide sequence of the mutant geneis disclosed in U.S. Pat. No. 4,769,061 to Comai. European PatentApplication Number 0 333 033 to Kumada, et al., and U.S. Pat. No.4,975,374 to Goodman, et al., disclose nucleotide sequences of glutaminesynthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EuropeanApplication Number 0 242 246 to Leemans, et al., De Greef, et al.,(1989) Bio/Technology 7:61, describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. See also, U.S. Pat. Nos. 5,969,213; 5,489,520;5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024;6,177,616 B1 and 5,879,903, which are incorporated herein by referencefor this purpose. Exemplary of genes conferring resistance to phenoxypropionic acids and cycloshexones, such as sethoxydim and haloxyfop, arethe Acc1-S1, Acc1-S2 and Acc1-S3 genes described by Marshall, et al.,(1992) Theor. Appl. Genet. 83:435. See also, U.S. Pat. Nos. 5,188,642;5,352,605; 5,530,196; 5,633,435; 5,717,084; 5,728,925; 5,804,425 andCanadian Patent Number 1,313,830, which are incorporated herein byreference for this purpose.

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla, et al.,(1991) Plant Cell 3:169, describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNumbers 53435, 67441 and 67442. Cloning and expression of DNA coding fora glutathione S-transferase is described by Hayes, et al., (1992)Biochem. J. 285:173.

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori, et al., (1995)Mol Gen Genet 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota, et al., (1994)Plant Physiol 106:17), genes for glutathione reductase and superoxidedismutase (Aono, et al., (1995) Plant Cell Physiol 36:1687, and genesfor various phosphotransferases (Datta, et al., (1992) Plant Mol Biol20:619).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1;and 5,767,373; and international publication WO 01/12825, which areincorporated herein by reference for this purpose.

3. Transgenes that confer or contribute to an altered graincharacteristic, such as:

(A) Altered fatty acids, for example, by

-   -   (1) Down-regulation of stearoyl-ACP desaturase to increase        stearic acid content of the plant. See, Knultzon, et al., (1992)        Proc. Natl. Acad. Sci. USA 89:2624 and WO99/64579 (Genes for        Desaturases to Alter Lipid Profiles in Corn),    -   (2) Elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-3 gene modification (see, U.S.        Pat. Nos. 6,063,947; 6,323,392; 6,372,965 and WO 93/11245),    -   (3) Altering conjugated linolenic or linoleic acid content, such        as in WO 01/12800,    -   (4) Altering LEC1, AGP, Dek1, Superal1, mi1ps, various Ipa genes        such as Ipa1, Ipa3, hpt or hggt. For example, see WO 02/42424,        WO 98/22604, WO 03/011015, U.S. Pat. Nos. 6,423,886, 6,197,561,        6,825,397, US Patent Application Publication Numbers        2003/0079247, 2003/0204870, WO02/057439, WO03/011015 and        Rivera-Madrid, et al., (1995) Proc. Natl. Acad. Sci.        92:5620-5624.

(B) Altered phosphate content, for example, by the

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant. For example, see, Van Hartingsveldt, et        al., (1993) Gene 127:87, for a disclosure of the nucleotide        sequence of an Aspergillus niger phytase gene.    -   (2) Up-regulation of a gene that reduces phytate content. In        maize, this, for example, could be accomplished, by cloning and        then re-introducing DNA associated with one or more of the        alleles, such as the LPA alleles, identified in maize mutants        characterized by low levels of phytic acid, such as in Raboy, et        al., (1990) Maydica 35:383 and/or by altering inositol kinase        activity as in WO 02/059324, US Patent Application Publication        Number 2003/0009011, WO 03/027243, US Patent Application        Publication Number 2003/0079247, WO 99/05298, U.S. Pat. Nos.        6,197,561, 6,291,224, 6,391,348, WO2002/059324, US Patent        Application Publication Number 2003/0079247, WO98/45448,        WO99/55882, WO01/04147.

(C) Altered carbohydrates effected, for example, by altering a gene foran enzyme that affects the branching pattern of starch, a gene alteringthioredoxin. (See, U.S. Pat. No. 6,531,648). See, Shiroza, et al.,(1988) J. Bacteriol 170:810 (nucleotide sequence of Streptococcus mutansfructosyltransferase gene), Steinmetz, et al., (1985) Mol. Gen. Genet.200:220 (nucleotide sequence of Bacillus subtilis levansucrase gene),Pen, et al., (1992) Bio/Technology 10:292 (production of transgenicplants that express Bacillus licheniformis alpha-amylase), Elliot, etal., (1993) Plant Molec Biol 21:515 (nucleotide sequences of tomatoinvertase genes), Søgaard, et al., (1993) J. Biol. Chem. 268:22480(site-directed mutagenesis of barley alpha-amylase gene) and Fisher, etal., (1993) Plant Physiol 102:1045 (maize endosperm starch branchingenzyme II), WO 99/10498 (improved digestibility and/or starch extractionthrough modification of UDP-D-xylose 4-epimerase, Fragile 1 and 2, Ref1,HCHL, C4H), U.S. Pat. No. 6,232,529 (method of producing high oil seedby modification of starch levels (AGP)). The fatty acid modificationgenes mentioned above may also be used to affect starch content and/orcomposition through the interrelationship of the starch and oilpathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683,US Patent Application Publication Number 2004/0034886 and WO 00/68393involving the manipulation of antioxidant levels through alteration of aphytl prenyl transferase (ppt), WO 03/082899 through alteration of ahomogentisate geranyl geranyl transferase (hggt).

(E) Altered essential seed amino acids. For example, see, U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO99/40209 (alteration of amino acid compositions inseeds), WO99/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO98/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO98/56935 (plant amino acid biosyntheticenzymes), WO98/45458 (engineered seed protein having higher percentageof essential amino acids), WO98/42831 (increased lysine), U.S. Pat. No.5,633,436 (increasing sulfur amino acid content), U.S. Pat. No.5,559,223 (synthetic storage proteins with defined structure containingprogrammable levels of essential amino acids for improvement of thenutritional value of plants), WO96/01905 (increased threonine),WO95/15392 (increased lysine), US Patent Application Publication Number2003/0163838, US Patent Application Publication Number 2003/0150014, USPatent Application Publication Number 2004/0068767, U.S. Pat. No.6,803,498, WO01/79516, and WO00/09706 (Ces A: cellulose synthase), U.S.Pat. No. 6,194,638 (hemicellulose), U.S. Pat. No. 6,399,859 and USPatent Application Publication Number 2004/0025203 (UDPGdH), U.S. Pat.No. 6,194,638 (RGP).

4. Genes that control pollination, hybrid seed production, ormale-sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar, et al., and chromosomal translocationsas described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen, et al., U.S. Pat. No. 5,432,068,describe a system of nuclear male sterility which includes: identifyinga gene which is critical to male fertility; silencing this native genewhich is critical to male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the barnase and the barstar gene (Paul, et al.,(1992) Plant Mol. Biol. 19:611-622).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014 and 6,265,640; all of which are hereby incorporatedby reference.

Also see, U.S. Pat. No. 5,426,041 (discovery relating to a method forthe preparation of a seed of a plant comprising crossing a male sterileplant and a second plant which is male fertile), U.S. Pat. No. 6,013,859(molecular methods of hybrid seed production) and U.S. Pat. No.6,037,523 (use of male tissue-preferred regulatory region in mediatingfertility), all of which are hereby incorporated by reference for thispurpose.

5. Genes that create a site for site specific DNA integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see, Lyznik, et al., (2003) “Site-Specific Recombinationfor Genetic Engineering in Plants”, Plant Cell Rep 21:925-932 and WO99/25821, which are hereby incorporated by reference. Other systems thatmay be used include the Gin recombinase of phage Mu (Maeser, et al.,1991), the Pin recombinase of E. coli (Enomoto, et al., 1983), and theR/RS system of the pSR1 plasmid (Araki, et al., 1992).

6. Genes that affect abiotic stress resistance (including but notlimited to flowering, ear and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress.

For example, see, WO 00/73475 where water use efficiency is alteredthrough alteration of malate; U.S. Pat. Nos. 5,892,009, 5,965,705,5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866, 6,717,034,6,801,104, WO2000060089, WO2001026459, WO2001035725, WO2001034726,WO2001035727, WO2001036444, WO2001036597, WO2001036598, WO2002015675,WO2002017430, WO2002077185, WO2002079403, WO2003013227, WO2003013228,WO2003014327, WO2004031349, WO2004076638, WO9809521 and WO9938977describing genes, including CBF genes and transcription factorseffective in mitigating the negative effects of freezing, high salinity,and drought on plants, as well as conferring other positive effects onplant phenotype; US Patent Application Publication Number 2004/0148654and WO01/36596 where abscisic acid is altered in plants resulting inimproved plant phenotype such as increased yield and/or increasedtolerance to abiotic stress; WO2000/006341, WO04/090143, U.S. patentapplication Ser. Nos. 10/817,483 and 09/545,334 where cytokininexpression is modified resulting in plants with increased stresstolerance, such as drought tolerance, and/or increased yield. Also seeWO0202776, WO03052063, JP2002281975, U.S. Pat. No. 6,084,153, WO0164898,U.S. Pat. No. 6,177,275 and U.S. Pat. No. 6,107,547 (enhancement ofnitrogen utilization and altered nitrogen responsiveness). For ethylenealteration, see, US Patent Application Publication Numbers 2004/0128719,2003/0166197 and WO200032761. For plant transcription factors ortranscriptional regulators of abiotic stress, see e.g., US PatentApplication Publication Number 2004/0098764 or US Patent ApplicationPublication Number 2004/0078852.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see, e.g.,WO97/49811 (LHY), WO98/56918 (ESD4), WO97/10339 and U.S. Pat. No.6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT), WO96/14414 (CON),WO96/38560, WO01/21822 (VRN1), WO00/44918 (VRN2), WO99/49064 (GI),WO00/46358 (FRI), WO97/29123, U.S. Pat. Nos. 6,794,560, 6,307,126 (GAI),WO99/09174 (D8 and Rht), and WO2004076638 and WO2004031349(transcription factors).

Seed Cleaning

This discovery is also directed to methods for producing cleaned canolaseed by cleaning seed of variety 45H33. “Cleaning a seed” or “seedcleaning” refers to the removal of foreign material from the surface ofthe seed. Foreign material to be removed from the surface of the seedincludes but is not limited to fungi, bacteria, insect material,including insect eggs, larvae, and parts thereof, and any other peststhat exist on the surface of the seed. The terms “cleaning a seed” or“seed cleaning” also refer to the removal of any debris or low quality,infested, or infected seeds and seeds of different species that areforeign to the sample. This discovery is also directed to producesubsequent generations of seed from seed of variety 45H33, harvestingthe subsequent generation of seed; and planting the subsequentgeneration of seed.

Seed Treatment

“Treating a seed” or “applying a treatment to a seed” refers to theapplication of a composition to a seed as a coating or otherwise. Thecomposition may be applied to the seed in a seed treatment at any timefrom harvesting of the seed to sowing of the seed. The composition maybe applied using methods including but not limited to mixing in acontainer, mechanical application, tumbling, spraying, misting, andimmersion. Thus, the composition may be applied as a slurry, a mist, ora soak. The composition to be used as a seed treatment can be apesticide, fungicide, insecticide, or antimicrobial. For a generaldiscussion of techniques used to apply fungicides to seeds, see “SeedTreatment,” 2d ed., (1986), edited by K. A Jeffs (chapter 9), hereinincorporated by reference in its entirety.

INDUSTRIAL APPLICABILITY

The seed of the 45H33 variety, the plant produced from such seed,various parts of the 45H33 hybrid canola plant or its progeny, a canolaplant produced from the crossing of the 45H33 variety, and the resultingseed, can be utilized in the production of an edible vegetable oil orother food products in accordance with known techniques. The remainingsolid meal component derived from seeds can be used as a nutritiouslivestock feed.

DEPOSIT

Applicant has made a deposit of at least 2500 seeds of canola variety45H33 with the American Type Culture Collection (ATCC), 10801 UniversityBoulevard, Manassas, Va. 20110-2209 USA, ATCC Deposit No. PTA-122029.The seeds deposited with the ATCC on Feb. 20, 2015 for PTA-122029 weretaken from the seed stock maintained by Pioneer Hi-Bred International,Inc., 7250 NW 62^(nd) Avenue, Johnston, Iowa 50131-1000 since prior tothe filing date of this application. Access to this deposit will beavailable during the pendency of the application to the Commissioner ofPatents and Trademarks and persons determined by the Commissioner to beentitled thereto upon request. Upon allowance of any claims in theapplication, the Applicant will make available to the public, pursuantto 37 C.F.R. §1.808, sample(s) of the deposit of at least 2500 seeds ofcanola variety 45H33 with the American Type Culture Collection (ATCC),10801 University Boulevard, Manassas, Va. 20110-2209. This deposit ofseed of canola variety 45H33 will be maintained in the ATCC depository,which is a public depository, for a period of 30 years, or 5 years afterthe most recent request, or for the enforceable life of the patent,whichever is longer, and will be replaced if it becomes nonviable duringthat period. Additionally, Applicant has satisfied all the requirementsof 37 C.F.R. §§1.801-1.809, including providing an indication of theviability of the sample upon deposit. Applicant has no authority towaive any restrictions imposed by law on the transfer of biologicalmaterial or its transportation in commerce. Applicant(s) do not waiveany infringement of their rights granted under this patent or rightsapplicable to canola hybrid 45H33 under the Plant Variety Protection Act(7 USC 2321 et seq.).

Varietal Characteristics (See Also Tables 1 Through 5)

Seed yield Similar yielding compared to the mean of the WCC/RRC checks;2.0% higher yielding than 45H29. Disease Classified as Resistant (R) toblackleg (Leptospaera reaction maculans) according to WCC/RRCguidelines. Classified as resistant (R) to Fusarium wilt and club root(Plasmodiophora brassicae) according to trials. Plant height Similarheight compared to the mean of the WCC/RRC checks. MaturityApproximately 0.5 days earlier maturing than the WCC/RRC checks. LodgingSimilar lodging rating to the mean of the WCC/RRC checks. HerbicideTolerant to glyphosate herbicides. Field testing tolerance confirms that45H33 tolerates the recommended rate of glyphosate (1.5 L/ha) withoutshowing plant injury or any significant negative effect on yield,agronomic, and quality traits. Variants Exhibits less than 1500/10,000(<15%) glyphosate susceptible plants.Seed Characteristics

Seed color Dark brown Grain size 1000 seed weight is similar to the meanof the WCC/RRC checks. Seed oil content 0.2% lower than the mean of theWCC/RRC checks. Seed protein content 0.6% higher than the mean of theWCC/RRC checks. Erucic acid Less than 0.5% (maximum allowable limit).Total saturates 0.17% higher than the mean of the WCC/RRC checks. Totalglucosinolates Canola quality - 0.5 umol/g lower than the WCC/RRCchecks. Chlorophyll Similar to the mean of the WCC/RRC checks.

TABLE 1 Variety Descriptions based on Morphological, Agronomic andQuality Traits 45H29 Trait 45H33 (Check Variety) Code Trait MeanDescription Mean Description 1.2 Seasonal Type Spring 2.1 Cotyledonwidth 5 Medium 5 Medium 3 = narrow 5 = medium 7 = wide 2.2 Seedlinggrowth 5 5 habit (leaf rosette) 1 = weak rosette 9 = strong rosette 2.3Stem anthocyanin 2 Absent or very 2 Absent or very intensity weak toweak weak to weak 1 = absent or very weak 3 = weak 5 = medium 7 = strong9 = very strong 2.4 Leaf type 1 Petiolate 8 1 = petiolate 9 = lyrate 2.5Leaf shape 3 Narrow elliptic 3 Narrow elliptic 3 = narrow elliptic 7 =orbicular 2.6 Leaf length 5 Medium 5 Medium 3 = short 5 = medium 7 =long 2.7 Leaf width 4 Narrow/Medium 4 Narrow/Medium 3 = narrow 5 =medium 7 = wide 2.8 Leaf color 2 Medium green 2.5 Medium green 1 = lightgreen to Dark green 2 = medium green 3 = dark green 4 = blue-green 2.12Leaf lobe 5 Medium 5 Medium development 1 = absent or very weak 3 = weak5 = medium 7 = strong 9 = very strong 2.13 Number of leaf 5 5 lobes 2.15Petiole length 5 Medium 3 Short 3 = short 5 = medium 7 = long 2.16 Leafmargin shape 3 Sharp 3 Sharp 1 = undulating 2 = rounded 3 = sharp 2.17Leaf margin 5 Medium 5 Medium indentation 1 = absent or very weak (veryshallow) 3 = weak (shallow) 5 = medium 7 = strong (deep) 9 = very strong(very deep) 2.18 Leaf attachment to 2 Partial clasping 2 Partialclasping stem 1 = complete clasping 2 = partial clasping 3 =non-clasping 3.1 Flower date 50% 46.1 3.2 Plant height at 7 Tallmaturity 3 = short 5 = medium 7 = tall 3.4 Flower bud location 1 Budsabove 1 Buds above 1 = buds above most most recently most recentlyrecently opened opened flowers opened flowers flowers 9 = buds belowmost recently opened flowers 3.5 Petal color 3 Medium yellow 3 Mediumyellow 1 = white 2 = light yellow 3 = medium yellow 4 = dark yellow 5 =orange 6 = other 3.6 Petal length 5 Medium 5 Medium 3 = short 5 = medium7 = long 3.7 Petal width 6 Medium/Wide 5 Medium 3 = narrow 5 = medium 7= wide 3.8 Petal spacing 6 Touching/Slight 6 Touching/Slight 1 = openoverlap overlap 3 = not touching 5 = touching 7 = slight overlap 9 =strongly overlap 3.11 Anther fertility 9 All anthers 9 All anthers 1 =sterile shedding pollen shedding pollen 9 = all anthers shedding pollen3.12 Pod (silique) length 5 Medium 5 Medium 1 = short (<7 cm) 5 = medium(7-10 cm) 9 = long (>10 cm) 3.13 Pod (silique) width 5 Medium (4 mm) 5Medium (4 mm) 3 = narrow (3 mm) 5 = medium (4 mm) 7 = wide (5 mm) 3.14Pod (silique) 3 Semi-erect 2 Erect/Semi- attitude erect 1 = erect 3 =semi-erect 5 = horizontal 7 = slightly drooping 9 = drooping 3.15 Pod(silique) beak 5 Medium 5 Medium length 3 = short 5 = medium 7 = long3.16 Pedicel length 5 Medium 4 Short/Medium 3 = short 5 = medium 7 =long 3.17 Maturity (days from 100.1 planting) 4.1 Seed coat color 1.5Black to brown 1.5 Black to brown 1 = black 2 = brown 3 = tan 4 = yellow5 = mixed 6 = other 4.3 Seed 3.6 weight/Thousand seed weight (5-6%moisture content): grams per 1,000 seeds 5.1 Shatter resistance 5.8 1 =Not tested 3 = Poor 5 = Fair 7 = Good 9 = Does not shatter 5.2 Lodgingresistance 6.3 Fair/Good 1 = not tested 3 = poor 5 = fair 7 = good 9 =excellent 6.3 Blackleg resistance 1 Resistant 0 = not tested 1 =resistant 3 = moderately resistant 5 = moderately susceptible 7 =susceptible 9 = highly susceptible 6.7 Fusarium wilt 1 Resistantresistance 0 = not tested 1 = resistant 3 = moderately resistant 5 =moderately susceptible 7 = susceptible 9 = highly susceptible 8.1 Oilcontent 47.51 percentage 8.5 Protein percentage 46.15 (whole dry seed)8.7 Glucosinolates 2 Low (10-15 2 Low (10-15 (μmoles total μmol pergram) μmol per gram) glucs/g whole seed) 1 = very low (<10) 2 = low(10-15) 3 = medium (15-20) 4 = high (>20) 8.8 Chlorophyll content 1 Low(<8 ppm) 1 Low (<8 ppm) (mg/kg seed) 1 = low (<8 ppm) 2 = medium (8-15ppm) 3 = high (>15 ppm)

Example 1 Herbicide Resistance

Appropriate field tests have shown that 45H33 tolerates the recommendedrate (1.5 L/ha) of glyphosate herbicide without showing plant injury orany significant negative effect on yield, agronomic, or quality traits.This hybrid exhibits less than 1500/10,000 (<15%) glyphosate-susceptibleplants.

TABLE 2 Effect of herbicide application on agronomic and quality traitsof 45H33 in herbicide tolerance trials in 2012 and 2013 % Stand DaysTreat- Yield Reduction to Height Days to % % Oil + Gluc's Variety mentq/ha (PCTSR) Flower (cm) Maturity Oil Protein Protein @ 8.5% Chlorophyll2012 Vegreville, AB 45H33 2X 17.0 0 51 95 100 50.0 44.5 94.5 17.0 0.045H29 2X 19.1 0 48 95 100 49.0 47.6 96.7 22.0 0.0 CV % 12.7 273.7 7.06.6 1.4 2.2 2.8 0.6 7.3 171.0 LSD (0.05) 2.8 0.7 4.9 8.8 1.9 2.3 2.7 1.23.0 2.0 SE 1.00 0.28 1.70 3.11 0.71 0.80 0.94 0.42 0.71 0.71 2012Saskatoon, SK 45H33 2X 18.8 1 119 102 46.9 46.6 93.5 15.0 4.0 45H29 2X15.9 0 118 103 48.3 47.3 95.6 17.0 6.0 CV % 12.7 109.8 10.5 0.7 1.5 1.40.8 7.3 38.8 LSD (0.05) 2.5 1.0 16.5 0.9 1.4 1.4 1.6 2.0 8.0 SE 0.900.35 5.87 0.35 0.50 0.47 0.57 0.71 2.83 2012 Average 45H33 2X 17.9 050.8 106.9 100.6 48.5 45.5 94.0 16.0 2.0 45H29 2X 17.5 0 47.5 106.3101.4 48.7 47.5 96.1 20.0 3.0 CV % 12.7 149.0 7.0 9.1 1.1 1.9 2.2 0.77.3 53.7 LSD (0.05) 3.3 0.7 4.9 10.4 1.3 1.7 1.8 1.5 2.0 6.0 SE 1.170.21 1.70 3.68 0.50 0.61 0.65 0.54 0.71 2.12 # 2 2 1 2 2 2 2 2 2 2Locations 2013 Vegreville, AB 45H33 2X 31.1 0 51 115 102 47.7 44.1 91.715.0 4.0 45H31 2X 27.8 0 50 115 102 49.9 44.1 93.9 11.0 3.0 CV % 8.3574.5 1.4 6.2 1.3 1.9 2.4 0.7 10.1 56.6 LSD (0.05) 3.9 1.0 1.0 11.0 2.01.6 1.8 1.1 2.0 4.0 SE 1.39 0.00 0.71 3.54 0.71 0.57 0.62 0.37 0.71 1.412013 Saskatoon, SK 45H33 2X 14.6 0 47 85 99 47.6 43.7 91.3 13.0 5.045H31 2X 15.3 0 46 83 99 48.9 44.7 93.6 13.0 4.0 CV % 15.0 296.6 1.710.7 1.2 1.4 1.9 0.6 7.7 54.0 LSD (0.05) 4.0 2.0 1.0 15.0 2.0 1.1 1.40.9 2.0 3.0 SE 1.42 0.71 0.71 5.66 0.71 0.40 0.49 0.31 0.71 1.41 2013Rosebank, MB 45H33 2X 33.5 0 40 120 94 47.3 46.6 93.9 18.0 0.0 45H31 2X30.9 0 40 115 95 47.8 48.5 96.3 16.0 0.0 CV % 7.9 160.6 1.5 7.6 1.0 1.61.1 0.5 3.4 110.8 LSD (0.05) 4.1 1.0 1.0 14.0 2.0 1.5 1.0 1.0 1.0 0.0 SE1.46 0.00 0.00 4.95 0.71 0.52 0.36 0.35 0.71 0.00 2013 Average 45H33 2X26.4 0 46.0 107.0 98.0 47.5 44.8 92.3 15.0 3.0 45H31 2X 24.6 0 46.0104.0 98.0 48.9 45.7 94.6 13.0 3.0 CV % 9.5 330.6 1.6 8.0 1.2 1.7 2.00.6 7.3 72.5 LSD (0.05) 2.4 1.0 1.0 9.0 1.0 0.9 1.1 0.9 1.0 2.0 SE 0.850.00 0.00 2.83 0.71 0.31 0.38 0.33 0.71 0.71 Locations 3 3 3 3 3 3 3 3 33 2 Year Average 45H33 2X 23.0 0 47 107 99 47.9 45.1 93.0 15.6 2.6 45H292X 17.5 0.3 47.5 106.3 101.4 48.7 47.5 96.1 20.0 3.0 45H31 2X 24.6 0.046.0 104.0 98.0 48.9 45.7 94.6 13.0 3.0 Avg. of 2X 21.8 0.0 46.0 105.0100.0 48.8 46.4 95.2 15.8 2.6 checks CV % 11.3 283.0 2.9 8.3 1.1 1.7 1.90.6 7.2 86.2 LSD (0.05) 1.4 0.5 1.8 6.2 0.7 0.6 0.7 0.4 0.8 1.6 SE 0.500.16 0.66 2.22 0.27 0.22 0.24 0.16 0.27 0.59 Locations 5 5 4 5 5 5 5 5 55

Example 2 Miscellaneous Disease Resistance

Blackleg

Blackleg tolerance was measured following the standard proceduredescribed in the Procedures of the Western Canada Canola/RapeseedRecommending Committee (WCC/RRC) Incorporated for the Evaluation andRecommendation for Registration of Canola/Rapeseed Candidate Cultivarsin Western Canada. Blackleg was rated on a scale of 0 to 5: a plant withzero rating is completely immune to disease while a plant with “5”rating is dead due to blackleg infection.

Canola variety “Westar” was included as an entry/control in eachblackleg trial. Tests are considered valid when the mean rating forWestar is greater than or equal to 2.6 and less than or equal to 4.5.(In years when there is poor disease development in Western Canada theWCC/RRC may accept the use of data from trials with a rating for Westarexceeding 2.0.)

The ratings are converted to a percentage severity index for each line,and the following scale is used to describe the level of resistance:

Classification Rating (% of Westar) R (Resistant) <30 MR (ModeratelyResistant) 30-49 MS (Moderately Susceptible) 50-69 S (Susceptible) 70-89HS (Highly Susceptible)  90-100

TABLE 3 Summary of Blackleg Ratings for 45H33 2012 Plum 2013 2 YearCoulee Alvena Boissevain Carman Vegreville Average % Westar Class 45H330.9 0.6 0.7 1.0 1.5 0.9 26.7 R Westar 3.8 3.3 2.9 3.4 3.8 3.4

Example 3 Summary of Performance of 45H33 in Two Years of Co-Op Testing

Two years (2012 and 2013) of trials were conducted. WCC/RRC guidelineswere followed for conducting trials. Each trial had four replicates andhad a plot size of 1.5 m×6 m. Yield and agronomic traits were recordedand seed samples were collected from two of the four replicates atselected sites and were analyzed for quality traits such as oil andprotein percent at 8.5% moisture, total whole seed glucosinolates at8.5%, chlorophyll, total saturated fatty acid, 1000 seed weight etc.WCC/RRC guidelines were followed for analyzing quality parameters.

TABLE 4 Summary of Performance of 45H33 in two years of Co-op TestingEarly Vigor Lodging Score Plant Oil % Yield Yield % Days to Days to (1 =poor, (1 = poor, Height @ zero Variety (q/ha) Check Maturity Flower 9 =best) 9 = best) (cm) moisture 2012 45H33 21.0 102 98.0 51.5 6.4 6.6 12747.0 5440 21.3 103 99.0 50.9 6.5 6.8 127 47.0 45H29 19.9 97 98.0 51.06.3 6.4 125 48.4 # Locs 16 16 14 10 21 9 14 10 Check 20.6 100 98.5 51.06.4 6.6 126 47.7 Avg. Diff. 0.4 2 −0.5 0.5 0.0 0.0 0.6 −0.7 from Check2013 45H33 38.3 99 99.7 42.7 6.2 124 47.9 5440 39.3 102 100.2 43.9 6.9124 47.0 45H29 37.8 98 99.9 43.2 5.9 124 48.5 # Locs 22 22 20 16 22 1914 Check 38.6 100 100.1 43.6 6.4 124 47.8 Avg. Diff. −0.2 −1 −0.4 −0.8−0.2 −0.3 0.1 from Check 2 Years Average 45H33 31.0 100 99.0 46.1 6.46.3 125 47.5 5440 31.7 103 99.7 46.6 6.5 6.9 125 47.0 45H29 30.3 97 99.146.2 6.3 6.0 125 48.4 # Locs 38 38 34 26 21 31 33 24 Check 31.0 100 99.446.4 6.4 6.5 125 47.7 Avg. Diff. 0.0 0.4 −0.4 −0.3 0.0 −0.1 0.1 −0.2from Check Total Shatter Score Protein % Total Glucs SaturatedChlorophyll Green 1000 Seed (1 = poor; Variety (oil free) (umol/g) Fat(%) (ppm) Seed % Weight (g) 9 = best) 2012 45H33 48.7 13.9 6.7 2.0 0.83.6 5.8 5440 47.3 12.2 6.4 2.0 0.3 3.9 5.8 45H29 48.2 14.2 6.6 2.0 0.53.5 5.7 # Locs 12 12 12 16 12 13 5 Check 47.7 13.2 6.5 2.0 0.4 3.7 5.8Avg. Diff. 1.0 0.7 0.16 0.0 0.4 −0.1 0.0 from Check 2013 45H33 43.9 11.66.9 5440 43.5 10.1 6.6 45H29 44.0 12.4 6.8 # Locs 14 14 14 Check 43.811.2 6.7 Avg. Diff. 0.2 0.4 0.18 from Check 2 Years Average 45H33 46.112.7 6.8 2.0 0.8 3.6 5.8 5440 45.2 11.1 6.5 2.0 0.3 3.9 5.8 45H29 45.913.2 6.7 2.0 0.5 3.5 5.7 # Locs 26 26 26 16 12 13 5 Check 45.6 12.1 6.62.0 0.4 3.7 5.8 Avg. Diff. 0.5 0.5 0.17 0.0 0.4 −0.1 0.0 from Check

Example 5 Clubroot Resistance in 45H33

Clubroot rating scale: Plants were scored on a 0-3 scale based on rootsymptoms

0=no galling

1=a few small galls (small galls on less than ⅓ of roots)

2=moderate galling (small to medium-sized galls on ⅓ to ⅔ of roots)

3=severe galling (medium to large-sized galls on more than ⅔ of roots)

Index of disease: The individual scores were used to calculate an indexof disease (ID)

${I\; D\mspace{14mu}(\%)} = {\frac{\sum\left( {{n \times 0} + {n \times 1} + {n \times 2} + {n \times 3}} \right)}{N \times 3} \times 100\%}$Where Σ is the sum total; n is the number of plants in a class; N is thetotal number of plants; and 0, 1, 2 and 3 are the symptom severityclasses.

TABLE 5 Clubroot disease Index for 45H33, 45H29 (R spring canola check)and susceptible commercial check 45H28 in 2011 field trials. Mean IDMean ID Field UofA GH UofA Mean ID % sus- Cat- Variety 2012 2013 meanceptible egory 45H33 9.8 9.0 9.4% 11% R 45H29 7.0 12.4 9.7% 12% RSusceptible 74.4 93.2 83.8%  100%  S 45H28 45H3145H33 (11N445R) is rated as resistant R for its reaction to Clubroot.

The foregoing discovery has been described in detail by way ofillustration and example for purposes of exemplification. However, itwill be apparent that changes and modifications such as single genemodifications and mutations, somaclonal variants, variant individualsselected from populations of the plants of the instant variety, and thelike, are considered to be within the scope of the present discovery.All references disclosed herein whether to journal, patents, publishedapplications and the like are hereby incorporated in their entirety byreference.

What is claimed is:
 1. A canola variety 45H33, representative seed ofthe variety having been deposited under ATCC Accession NumberPTA-122029.
 2. A seed of the canola variety of claim
 1. 3. The seed ofclaim 2, further comprising a seed treatment.
 4. The seed of claim 3,wherein the treatment comprises a fungicide or insecticide.
 5. A methodcomprising cleaning the seed of claim
 2. 6. A method of producing acanola seed, the method comprising planting the seed of claim 2 toproduce a subsequent generation of seed; harvesting the subsequentgeneration of seed; and planting the subsequent generation of seed. 7.The seed of claim 2, further comprising a trait introduced bybackcrossing or genetic transformation.
 8. The seed of claim 7, whereinthe trait is selected from the group consisting of male sterility, asite for site-specific recombination, abiotic stress tolerance, alteredphosphate, altered antioxidants, altered fatty acids, altered essentialamino acids, altered carbohydrates, herbicide resistance, insectresistance and disease resistance.
 9. The seed of claim 7, furthercomprising a seed treatment.
 10. The seed of claim 9, wherein thetreatment comprises a fungicide or insecticide.
 11. A method comprisingcleaning the seed of claim
 7. 12. A plant or plant part of the canolavariety of claim
 1. 13. A method of producing a second canola plant, themethod comprising applying plant breeding techniques to the plant orplant part of claim 12, thereby producing the second canola plant.
 14. Amethod for producing a canola plant derived from canola variety 45H33,the method comprising: (a) crossing the plant of claim 12 with itself ora second plant to produce progeny seed; (b) growing the progeny seed toproduce a progeny plant and crossing the progeny plant with itself or adifferent plant to produce further progeny seed; and (c) repeating step(b) for at least one additional generation to produce a canola plantderived from the variety 45H33.
 15. A method for producing a secondcanola plant or plant part, the method comprising doubling haploid seedgenerated from the plant of claim 12, thereby producing the secondcanola plant or plant part.
 16. The plant or plant part of claim 12,further comprising a trait introduced by backcrossing or genetictransformation.
 17. The plant or plant part of claim 16, wherein thetrait is selected from the group consisting of male sterility, a sitefor site-specific recombination, abiotic stress tolerance, alteredphosphate, altered antioxidants, altered fatty acids, altered essentialamino acids, altered carbohydrates, herbicide resistance, insectresistance and disease resistance.
 18. A method for producing a secondcanola plant or plant part, the method comprising applying plantbreeding techniques to the plant or plant part of claim 16, therebyproducing the second canola plant.
 19. A method for producing a canolaplant derived from canola variety 45H33, the method comprising: (a)crossing the plant of claim 16 with itself or a second plant to produceprogeny seed; (b) growing the progeny seed to produce a progeny plantand crossing the progeny plant with itself or a different plant toproduce further progeny seed; and (c) repeating step (b) for at leastone additional generation to produce a canola plant derived from thevariety 45H33 and further comprising the trait.
 20. A method forproducing a second canola plant or plant part, the method comprisingdoubling haploid seed generated from the plant of claim 16, therebyproducing the second canola plant or plant part.